[Correlation between fluid parameter and cardiovascular events in hemodialysis patients based on repeated body composition analyses].

Objective: To explore the correlation between fluid load index and cardiovascular events in hemodialysis patients based on repeated body composition analyses. Methods: A prospective cohort study was conducted to collect the clinical data of patients undergoing maintenance hemodialysis (MHD) in the Department of Nephrology, Hangzhou Hospital of Traditional Chinese Medicine from July to September 2020. The pre-dialysis fluid overload (FO) index-overhydration (OH) was measured by bioelectrical impedance method, and the follow-up monitoring was conducted every 6 months. According to the baseline OH value, patients were divided into non-FO group (OH≤2.5 L) and FO group (OH>2.5 L). Moreover, according to the repeated measurements, the patients were divided into continuous non-FO group, continuous FO group and intermittent FO group. All patients were followed up until October 1, 2022, and the outcomes were recorded. The end point of follow-up was cardiovascular events. The cumulative incidence of cardiovascular events was calculated by the Kaplan-Meier method, and the risk factors of cardiovascular events were analyzed by Cox proportional hazards regression model. Results: A total of 289 patients were included, including 88 patients (30.4%) with FO and 201 patients (69.6%) with non-FO. There were 168 males (58.1%) and 121 (41.9%) females, with an average age of (58±13) years and an average follow-up time of (22.0±6.5) months. Kaplan-Meier analysis showed that the incidence of cardiovascular events in the baseline FO group was higher than that in the non-FO group (log-rank χ2=14.44, P<0.001). The incidence of cardiovascular events in both the continuous FO group and the intermittent FO group was higher than that in the continuous non-FO group (log-rank χ2=41.47, P<0.001; log-rank χ2=18.36, P<0.001). After adjustment for gender, age, comorbidities, and biochemical indicators, the incidence of cardiovascular events in the baseline FO group was 1.850 times of the non-FO group (95%CI: 1.046-3.271, P=0.034). The incidence of cardiovascular events in the continuous FO group was 4.679 times of the continuous non-FO group (95%CI: 2.189-10.002, P<0.001). The incidence of cardiovascular events in the intermittent FO group was 3.410 times of the continuous non-FO group (95%CI: 1.696-6.857, P=0.001). Conclusions: OH value measured by bioelectrical impedance can be used as an important reference index for clinical monitoring of cardiovascular events in MHD patients. Continuous chronic and intermittent exposures to FO are risk factors for cardiovascular events in hemodialysis patients.

Direct Measurement of the Astrophysical

Fluorine is one of the most interesting elements in nuclear astrophysics, where the ^{19}F(p,α)^{16}O reaction is of crucial importance for Galactic ^{19}F abundances and CNO cycle loss in first generation Population III stars. As a day-one campaign at the Jinping Underground Nuclear Astrophysics experimental facility, we report direct measurements of the essential ^{19}F(p,αγ)^{16}O reaction channel. The γ-ray yields were measured over E_{c.m.}=72.4-344 keV, covering the Gamow window; our energy of 72.4 keV is unprecedentedly low, reported here for the first time. The experiment was performed under the extremely low cosmic-ray-induced background environment of the China JinPing Underground Laboratory, one of the deepest underground laboratories in the world. The present low-energy S factors deviate significantly from previous theoretical predictions, and the uncertainties are significantly reduced. The thermonuclear ^{19}F(p,αγ)^{16}O reaction rate has been determined directly at the relevant astrophysical energies.

[Three-dimensional movement of posterior teeth after losing the interproximal and occlusal contacts in adults].

OBJECTIVE: To observe the movement of posterior teeth after losing the interproximal and occlusal contacts in adults from the amount and speed of mesial-distal, buccal-lingual, occlusal-gingival and three-dimensional movements. METHODS: Twenty cases of metal post-core restoration from the Department of Prosthodontics, Peking University School and Hospital of Stomatology were recruited into this study, which was an observational study. The restored teeth had complete mesial, distal, and occlusal contacts before treatment. All the interproximal and occlusal contacts were removed during the preparation for the post-core. The three-dimensional positions of the post-core preparation were obtained by the intraoral scanner at the day of tooth preparation and that of post-core placement. The amounts of mesial-distal, buccal-lingual and occlusal-gingival tooth movement were measured in the software. On this basis, the amount of three-dimensional tooth movement was calculated. The speed of tooth movement was calculated based on the elapsed time between the two scans. RESULTS: Ten females and ten males with an average age of (29.5±4.9) years were recruited. The average elapsed time was (10.9±2.7) days. The amount of the mesial-distal tooth movement was (134.8±61.2) µm, of the buccal-lingual tooth movement was (110.3±39.5) µm, of the occlusal-gingival tooth movement was (104.8±57.5) µm, and of the three-dimensional tooth movement was (211.4±71.0) µm, respectively. The amounts of mesial-distal, buccal-lingual and three-dimensional tooth movements were larger in female than in male (P < 0.05). The speed of the mesial-distal tooth movement was (13.1±7.8) µm/d, of the buccal-lingual tooth movement was (10.6±4.5) µm/d, of the occlusal-gingival tooth movement was (10.1±6.8) µm/d, and of the three-dimensional tooth movement was (20.5±9.7) µm/d, respectively. The speed of mesial-distal and buccal-lingual tooth movements were larger in female than in male (P < 0.05). The speed of three-dimensional tooth movement was slightly larger in female than in male, while there was no significant difference between different genders (P>0.05). CONCLUSION: The three-dimensional position of posterior teeth changed after losing the interproximal and occlusal contacts in adults. The female had more significant and faster tooth movement than the male.

[Blood pressure and heart rate changes of 640 single dental implant surgeries].

OBJECTIVE: To investigate the blood pressure and heart rate changes and influencing factors during single dental implant surgery. METHODS: A retrospective cohort study was conducted. Six hundred and forty cases underwent single dental implant placement in Department of Prosthodontics, Peking University School and Hospital of Stomatology from January 2016 to December 2016 were recruited in this study according to the inclusion and exclusion criteria. They were divided into different groups according to the exposure factors which were patient characteristics (gender, age) and surgical procedures (immediate placement, flap elevation, bone grafting). The correlation between blood pressure and heart rate variability during single dental implant surgery and the patient characteristics and surgical procedures were analyzed. RESULTS: The average systolic blood pressure variability was 9.47%±6.45% (maximum 46.04%), the average diastolic blood pressure variability was 12.18%±9.39% (maximum 88.00%), and the average heart rate variability was 10.59%±7.68% (maximum 49.12%). The effects of age and bone grafting on blood pressure variability, and of gender and immediate placement on heart rate variability, were statistically significant (P < 0.05), respectively. The incidence of abnormal intraoperative blood pressure rise was 4.69%, of which there were 4 cases (0.63%) of systolic blood pressure ≥180 mmHg and/or diastolic blood pressure ≥110 mmHg. The incidence of abnormal intraoperative heart rate rise was 6.72%. Hierarchical analysis showed a higher risk of abnormal intraoperative blood pressure rise in the elderly (≥60 years) male patients [P < 0.05, RR=3.409 (95%CI: 1.155-10.062) ] and flap elevation with bone grafting cases [P < 0.05, RR=2.382 (95%CI: 1.126-5.040)], respectively. There was no statistically significant association between abnormal heart rate rise and patient characteristics or surgical procedures (P>0.05). CONCLUSION: There was a certain risk of blood pressure and heart rate variability during dental implant surgery. Elderly male patients and flap elevation with bone grafting were risk factors of abnormal intraoperative blood pressure rise.

[Effects of loupes and microscope on the prosthodontist's posture from ergonomic aspects].

OBJECTIVE: To assess the effects of loupes and microscope on the posture of prosthodontists when preparing the laminate veneer, and to assess the clinical value of loupes and microscope from the ergonomic aspects. METHODS: Twenty young prosthodontists from Department of Prosthodontics, Peking University School and Hospital of Stomatology were recruited into this study, which was a prospective, single blind, self-control trials. The research hypothesis was concealed and the participants were deceived about the precise purpose of the study to counterbalance the lack of direct blinding. The prosthodontists prepared laminate veneers of open window type in the artificial dental model, under routine visual field (control group), 2.5× headwear loupes (loupes group), and 8× operating microscope (microscopic group) by turning. The participants were photographed from profile view and front view. Thereafter, the subjective assessment was performed by themselves using the visual analogue score (VAS). The expert assessment was performed by two professors using modified-dental operator posture assessment instrument on the basis of photographs of the profile view and front view. RESULTS: The subjective assessment scores for the control group, loupes group and microscopic group were 4.55±1.96, 7.90±1.12, and 9.00±0.92, respectively. There was significant difference between the three groups' subjective scores (P < 0.05). The expert assessment scores for the control group, loupes group and microscopic group were 16.38±1.52, 15.15±1.30, and 13.60±0.88, respectively. There was significant difference between the three groups' expert assessment scores (P < 0.05). Specifically, the three groups' expert assessment scores were significantly different (P < 0.05) in trunk position (front to back) (1.33±0.41, 1.03±0.11, 1.00±0.00), head and neck position (front to back) (2.75±0.38, 2.13±0.36, 1.23±0.38), elbows level (1.38±0.43, 1.40±0.45, 1.13±0.22), and shoulders level (1.43±0.41, 1.23±0.34, 1.13±0.28). Thereinto, the microscopic group was better than loupes group in head and neck position (front to back) and elbows level (P < 0.05). CONCLUSION: Loupes and microscope improve the posture of the prosthodontist when preparing the laminate veneer, in which the microscope is better than loupes. Therefore, the magnification devices have clinical value from the ergonomic aspects.

[Effects of different surface treatments on the zirconia-resin cement bond strength].

OBJECTIVE: To evaluate the effects of different surface treatments on the shear bond strength between zirconia and resin cement. METHODS: Forty zirconia discs were randomly divided into four groups (10 discs in each group) for different surface treatments: control, no surface treatment; sandblast, applied air abrasion with aluminum oxide particles; ultraviolet (UV), the zirconia sample was placed in the UV sterilizer at the bottom of the UV lamp at 10 mm, and irradiated for 48 h; cold plasma, the discs were put in the cold plasma cabinet with the cold plasma generated from the gas of He for 30 s. Specimens of all the groups were surface treated prior to cementation with Panavia F 2.0 cement. The surface morphology and contact angle of water were measured. The shear bond strengths were tested and the failure modes were examined with a stereomicroscope. RESULTS: Surface morphology showed no difference between the UV/cold plasma group and the control group. Sandblasted zirconia displayed an overall heterogeneous distribution of micropores. The contact angle of the control group was 64.1°±2.0°. After sandblasting, UV irradiation and cold plasma exposure, the values significantly decreased to 48.8°±2.6°, 27.1°±3.6° and 32.0°±3.3°. The values of shear bond strength of the specimens with sandblasted (14.82±2.01) MPa were higher than those with no treatment (9.41±1.07) MPa with statistically significant difference (P<0.05). The values of shear bond strength of the specimens with UV irradiation (10.02±0.64) MPa were higher than those with no treatment (9.41±1.07) MPa, but without statistically significant difference (P>0.05). The values of cold plasma group (18.34±3.05) MPa were significantly higher than those of control group (9.41±1.07) MPa, even more than those with sandblast(14.82±2.01) MPa (P<0.05). X-ray photoelectron spectroscopy (XPS) showed increase in oxygen (O) and decrease in carbon (C) elements after UV and cold plasma treatment. The surface C/O ratio also decreased after UV and cold plasma treatment. CONCLUSION: Zirconia specimens treated with UV and cold plasma could significantly improve the hydrophilicity. The surface morphology was unaffected by the UV irradiation and cold plasma treatments. The improvements of ziconia shear bond strength were slight in UV group without statistically significant difference. Cold plasma treatment significantly improved the shear bond strength between zirconia and resin cement.

[A prospective multi-center trial of non-interventional and observational study of lenalidomide in Chinese patients with multiple myeloma].

Objective: To evaluate the efficacy and safety of lenalidomide in a real-world clinical practice in Chinese patients with multiple myeloma (MM). Methods: It was a prospective, multi-center, observational study. A total of 165 consecutive patients with MM treated with lenalidomide-based regimens were enrolled in 12 hospitals from June 2013 to November 2015. Relevant information was recorded, such as baseline clinical data, cytogenetic abnormalities, treatment regimens, and duration of treatment, safety, and survival. Results: (1)There were 126 relapsed and refractory MM (RRMM) patients, 25 newly diagnosed patients and 19 maintenance patients. The evaluable RRMM patients accounted for 120 cases, among which 74 cases(61.7%) reached the partial response (PR) or above, and a very good partial response (VGPR) in 16 patients (13.3%), a complete response (CR) in 14 cases (11.7%), a strictly complete response (sCR) in 4 cases (3.3%). Thus, a VGPR or above in 34 patients accounted for 28.3%. (2)The median follow-up was 13 months, the median time to progression 12 months. The median survival after receiving lenalidomide was 19 months, and the median overall survival (OS) was 62 months. (3) The univariate analysis in 120 RRMM patients suggested that prognostic factors for significant improvement in PFS included normal karyotype, international staging system (ISS) Ⅰ-Ⅱ, t(4; 14) negative (detected by fluorescence in situ hybridization), non-bortezomib resistance and response to previous regimens. As to OS, non-bortezomib resistance, response to previous regimens and non-primary refractoriness were positive factors. Multivariate analysis showed that the response to previous regimens (PR or better) was an independent good prognostic factor for progress-free survival(PFS), non-bortezomib resistance and non-primary refractoriness for OS. (4) Grade 3 or 4 adverse events that occurred in more than 10% of all enrolled patients were neutropenia (12.7%), leukocytosis(11.5%) and thrombocytopenia (12.7%). Owing to intolerance of toxic side effects, 7 cases withdrew lenalidomide. Conclusions: No matter what combination, regimens containing lenalidomide are effective to RRMM patients with overall response rate 61.7%, a time to progression 12 months and an overall survival 62 months.The toxicity is quite tolerable and manageable. In addition, the response to previous treatment (reached PR or above) is the independent good prognostic factor for PFS, non-bortezomib resistance and non-primary refractoriness for OS. Clinical trail registration: Clinicaltrials.gov, NCT01947309.

Biochemical characterization and functional analysis of UDP-glucose dehydrogenase, in the synthesis of biopolymer Ss from Sphingomonas sanxanigenens NX02.

Biopolymer Ss of Sphingomonas sanxanigenens strain NX02 is an sphingan that can be extracted using a small quantity of acid, which is a low cost extraction process. A UDP-glucose dehydrogenase gene (ugdG), related to Ss biosynthesis, was cloned from S. sanxanigenens NX02 and expressed in Escherichia coli. It encoded a 454-residue protein of 48.2 kDa. The deduced amino acid sequence had 77% identity with UDP-glucose dehydrogenase (UgdG) from Sphingomonas sp. KC8, and 73% identity with UgdG from Sphingomonas elodea ATCC31461. Purified recombinant UgdG had maximum activity at 35°C and pH 8.0, with Km values of 0.47 and 0.38 mM for UDP-glucose and NAD+, respectively. Overexpression of the ugdG gene in S. sanxanigenens resulted in increased (14.9 ± 0.5)% Ss production and higher fermentation broth viscosity. Furthermore, the weight-average molecular weight of polymer Ss from the recombinant strain was (5.3 ± 0.16)% higher and the viscosity was (74 ± 0.15)% higher than those from the WT strain at a shear rate of 1 rev/min.

First Report of Actinidia virus A and Actinidia virus B on Kiwifruit in China.

At present, two viruses affecting kiwifruit (Actinidia spp.), Actinidia virus A (AcVA) and Actinidia virus B (AcVB), both belonging to the genus Vitivirus in the family Betaflexiviridae, have been reported from New Zealand (2). The infected trees showed leaf vein chlorosis, flecking, and ringspots. China is the largest commercial kiwifruit producer. During field investigations in the growing season of 2013, symptoms of leaf chlorosis or ringspots, similar to those caused by AcVA and AcVB (1), were observed on some kiwifruit (Actinidia chinensis) plants in Hubei Province in the central China. Leaf samples were collected from three symptomatic and two symptomless plants of two A. chinensis cultivars. Total nucleic acids were extracted from the samples using a CTAB-based protocol described by Li et al. (3) and used as template in RT-PCR for the detection of AcVA and AcVB. Each virus was detected using two sets of primers reported by Blouin et al. (1). Primer sets AcVA 1F/1R and AcVA5F/5R were used for the AcVA detection, and AcVB1F/1R and AcVB5F/Viti3'R were used for the AcVB detection. AcVA was detected in three symptomatic plants (ID: Ac-HN-1, Ac-HN-3, and Ac-HN-5), and AcVB was detected in two symptomatic plants (ID: Ac-HN-1 and Ac-HN-3) and in one symptomless plant (ID: Ac-HN-2). Neither virus was detected in the second symptomless plant (ID: Ac-HN-4). Samples Ac-HN-1 and Ac-HN-3 had mixed infection of AcVA and AcVB, and sample Ac-HN-2 had the latent infection of AcVB. The sequenced 283-bp RT-PCR amplicons of the replicase-encoding gene from AcVA isolates AC-HN-3 and AC-HN-5 using AcVA1F/1R shared 90.8% nucleotide (nt) identity with the corresponding sequence of the New Zealand AcVA isolate (GenBank Accession No. JN427014.1). The 269-bp fragments of the RNA-binding protein-encoding gene obtained by using AcVA5F/5R shared 85.5 to 85.9% nt identities with the corresponding sequence of JN427014.1. The AcVB5F/Viti3'R products of 365 to 369 bp from three AcVB isolates shared 85.5 to 88.6% nt identities with the corresponding sequence of the New Zealand AcVB isolate. The representative sequences were submitted to GenBank with accession numbers KJ696776 and KJ696777 for the 269-bp fragments of AcVA-HN-1 and AcVA-HN-3, and KJ696778 and KJ696779 for the 365-bp and 369-bp fragments of AcVB-HN-1 and AcVB-HN-2, respectively. In addition, 12 and 14 out of 42 kiwi samples (excluding HN-1 to HN-5) collected randomly were positive for AcVA and AcVB as detected by RT-PCR. Meanwhile, the sample affected by AcVA-HN-5 was subjected to deep sequencing of the small RNAs (sRNAs) for complete survey of the infecting viruses. De novo assembly of sRNAs generated four sequence contigs, with lengths ranging from 161 to 285 nt, matching to ORFs 1 to 3 of the genome of the New Zealand AcVA isolate with significant nucleotide (91 to 95%) and amino acid (80 to 94%) similarities, and some other contigs from a new virus (unpublished). The result further confirmed AcVA infection in the kiwi plant. To our knowledge, this is the first report of both AcVA and AcVB outside of New Zealand. The Chinese isolates of the two viruses are distinct from those reported from New Zealand. The results provide valuable information for improving the viral sanitary status of the kiwifruit germplasm in China. References: (1) A. G. Blouin et al. Arch. Virol. 157:713, 2012. (2) A. G. Blouin et al. J. Plant Pathol. 95:221, 2013. (3) R. Li et al. J. Virol. Methods 154:48, 2008.

Effect of Sc contents on the mechanical properties and damping capacity of Al-Zn-Mg-Cu-Zr-Sc alloys with different grain structures.

Al-Zn-Mg-Cu-Zr-Sc alloys with different Sc contents were fabricated by casting, deformation, and T6 treatment. Deformation methods including rolling and friction-stir processing (FSP) were used to design their grain structure. A low additive amount (0.1) of Sc cannot refine the grains of the alloy with rolling and T6 treatment, and it instead coarsened the grains. The reason was the non-uniform distribution of nanosize Al3(Sc,Zr) phases that led to the occurrence of abnormal grain growth during homogenization. Meanwhile, the alloy with only 0.1Sc exhibited finer grains after FSP and T6 treatment than the alloys subjected to the same process but with higher Sc additive amount. Alloys with rolling-induced elongated grain structure exhibited better mechanical properties, and alloys with FSP-induced fine equiaxed grain structure exhibited higher high-strain and high-temperature internal friction values. These features are important performance parameters for applications in fields where vibration and noise are sensitive. The optimum additive amounts of Sc for alloys with elongated and fine equiaxed grain structures were 0.25 and 0.1, respectively.

First Report of Cherry necrotic rusty mottle virus on Stone Fruit Trees in China.

During the growing seasons of 2010 through 2012, leaf tissues from 206 stone fruit trees, including one flowering cherry, three sour cherry, six nectarine (Prunus persica L. var. nucipersica Schneider), 14 apricot, 24 plum (P. domestica L.), 41 sweet cherry, and 117 peach [P. persica (L.) Batsch] trees, grown in six provinces of China, were randomly collected and tested for the CNRMV infection by RT-PCR. Out of those sampled trees, 37 showed shot holes and vein yellowing symptoms. Total RNA was extracted from leaves using the CTAB protocol reported by Li et al. (2). The primer pair CGRMV1/CGRMV2 (1) was used to amplify a fragment of 949 bp from CNRMV genome, which includes the CP gene (804 bp). PCR products with the expected size were detected in one sweet cherry, one apricot, one peach, one plum, and two sour cherry plants. However, no correlation between PCR data and symptom expression could be found. PCR products were cloned into the vector pMD18-T (TaKaRa, Dalian, China). Three independent clones from each isolate were sequenced by Genscript Corp., Nanjing, China, and sequences were deposited in the GenBank under accession nos. JX491635, JX491636, JX491637, JX648205, and JX648206. Results of sequence analysis showed that sequences of the five CNRMV isolates shared the highest nt (99.0 to 99.6%) and aa (98.9 to 100%) similarities with a cherry isolate from Germany (GenBank Accession No. AF237816). The sequence of one isolate from a peach tree (JX648205) was divergent and shared only 84.7 to 86.1% nt and 94.4 to 95.1% aa similarities with those cp sequences. Clones intra each isolate shared more than 99% nt similarities. To confirm CNRMV infection, seedlings of peach GF 305 were graft-inoculated with bud-woods from a peach and a sweet cherry tree, which was positive to CNRMV and also two other viruses: Cherry green ring mottle virus (CGRMV) and Plum bark necrosis stem pitting-associated virus (PBNSPaV), as tested by RT-PCR. Grafted seedlings were kept in an insectproof greenhouse and observed for symptom development. In May of the following year, some newly developed leaves of inoculated seedlings showed vein yellowing, ringspot, and shot hole symptoms. Results of Protein A sandwich (PAS)-ELISA using an antiserum raised against the recombinant CP of a CNRMV isolate (unpublished) and RT-PCR confirmed CNRMV infection in inoculated trees. In addition to CNRMV, tested seedlings were also found to be infected with CGRMV and PBNSPaV by RT-PCR. To our knowledge, this is the first report on the occurrence of CNRMV on stone fruit trees in China, and also the first record of the CNRMV infection in peach and plum plants. Given the economic importance of its hosts and the visible symptoms of the viral disease, it is important to prevent the virus spread by using virus-tested propagation materials. References: (1) R. Li and R. Mock. J. Virol. Methods 129:162, 2005. (2) R. Li et al. J. Virol. Methods 154:48, 2008.

Mesenchymal stem cells differentially mediate regulatory T cells and conventional effector T cells to protect fully allogeneic islet grafts in mice.

AIMS/HYPOTHESIS: Limited information is available on the cellular interactions between regulatory T (T(reg)) cells and mesenchymal stem cells (MSCs). In particular, a direct effect of MSCs on the survival and proliferation of T(reg) cells has not been demonstrated. METHODS: We investigated the effects of MSCs on effector T (T(eff)) cells and T(reg) cells, and the molecular mechanisms involved in the distinct regulation of these two cell populations by MSCs in vivo and in vitro. RESULTS: We show that MSCs are capable of selectively suppressing T(eff) cells and fostering the generation of T(reg) cells. T(eff) cells, but not T(reg) cells, fail to respond to IL-2 and undergo profound apoptosis in the presence of MSCs. The differential regulations of these two T cell subsets by MSCs are associated with their distinct expressions of CD25, with MSCs specifically reducing the expression of CD25 on T(eff) and sparing T(reg) cells intact. In vivo, the administration of MSCs significantly delays the rejection of allogeneic islet grafts in adaptive transferred recipients by favouring the induction of T(reg) cells. In this model, MSCs inhibit the proliferation and development of alloreactive T(eff) but potently enhance the induction of T(reg) cells. CONCLUSIONS/INTERPRETATION: We demonstrate that MSCs are capable of regulating T(eff) and T(reg) cells differentially in vitro. MSCs inhibit T(eff) cells by inducing apoptosis and impairing the proliferative response to IL-2 in T(eff) cells, but favour the survival and expansion of T(reg) cells. This result is further demonstrated in mice that have undergone allogeneic islet transplantation, in which MSCs suppress alloreactive T(eff) cells while favouring the induction of T(reg) cells, thus protecting the islet allografts from rejection.

Scaling the dynamic electron scattering in imaging the graphene sheets by the high-angle annular dark-field microscopy.

Employing the graphene sheets (GSs), the electron scattering constants are measured in the high-angle annular dark-field (HAADF) imaging by the scanning transmission electron microscopy. Single scattering is found to be dominant until the layer number of 200, complying with a simple relation of I = Io(1 - e(-tau/lambda)). The discrete layer counting of the GSs enables precise determination of incident depths. This work results values of lambda = 48.2, 61.4, 97.9 and 115.6 nm for 80, 120, 160 and 200 keV electrons, respectively. The uncertainties with the mean free paths and the cross sections are confined to 10 percent. The dependences on the electron beam energy and the collection angle are discussed based on a multislice simulation.

Effects of enrofloxacin on cytochromes P4501A and P4503A in Carassius auratus gibelio (crucian carp).

Currently, although enrofloxacin (EF) as a widely used veterinary medicine has begun to apply to treating fish bacterial infections, the researches on the effects of EF on their main drug metabolic enzymes are limited. To investigate the effects of EF on fish cytochromes P450 (CYPs) 1A and 3A, the enzymatic activities and expressions (mRNA and protein) of crucian carp CYP1A and CYP3A after EF administration were examined. For CYP1A, in the in vivo experiments, EF exhibited potent inhibition on the CYP1A-related ethoxyresorufin-O-deethylase (EROD) activity, as well as CYP1A expressions at both protein and mRNA levels, at 24 h after administration with different EF dosages (3, 10, 30, and 60 mg/kg); Furthermore, CYP1A enzymatic activity and expressions at both protein and mRNA levels decreased more with increasing EF dosages. Additionally, the in vitro experimental results showed that, after incubated with microsomes, EF did not change the EROD activity through interacting directly with CYP1A. For CYP3A, the in vitro and in vivo experimental results demonstrated that EF could inhibit the CYP3A-related erythromycin N-demethylase activity in a time- and dose-dependent manner, while it did not suppress CYP3A expressions at both protein and mRNA levels after administration with EF for a short period (no more than 24 h); however, after injection with EF at a high dose (10 mg/kg) for a long period, the CYP3A protein and mRNA reached their lowest levels at 96 and 48 h, respectively. These results indicate that EF can suppress CYP1A expressions in a dose-dependent manner, thereby inhibiting further its catalytic activity; meanwhile, both the interactions of EF with CYP3A and the expressions decrease (protein and mRNA) caused by EF contribute to the CYP3A inhibition.

Efficacy and safety of itraconazole pulses vs. continuous regimen in cutaneous sporotrichosis.

BACKGROUND: Primary observation of using itraconazole pulse regimen (commonly used for onychomycosis) in five sporotrichosis patients produced satisfactory effect, but there are no randomized controlled trials to assess this regimen and to compare with routine continuous regimen. OBJECTIVES: The aim of this study was to compare the efficacy and safety of itraconazole pulse regimen, as a new treatment option for cutaneous sporotrichosis, with continuous regimen. METHODS: A prospective, randomized, evaluator-blind, controlled study was performed. A total of 50 cutaneous sporotrichosis patients were divided randomly into two groups: Pulsed Rx (npulse=25), treated with itraconazole 200 mg b.i.d. for 1 week and off for 3 weeks; Daily Rx (ncont=25), treated with itraconazole 100 mg bid continuously. In total, 46 patients (npulse=22, ncont=24) were assessable at the end of the study. Cure rates at weeks 12, 24 and 48 along with the course of treatment and the rate of side effects were evaluated. RESULTS: In Pulsed Rx, the cure rates at weeks 12, 24 and 48 were 77.3%, 81.8%, 81.8% respectively. The mean course of treatment was 2.65±0.81 pulses and the rate of side effects was 4.5%; In Daily Rx, the corresponding cure rates were 79.2%, 91.7% and 95.8% respectively. The mean course of treatment was 2.80±2.33 months and the rate of side effects was 16.7%. These parameters had no significant difference between the two groups (P>0.05). CONCLUSION: The efficacy and safety of Itraconazole pulse regimen for cutaneous sporotrichosis were similar to the continuous regimen. It is an effective and safe alternative treatment for cutaneous sporotrichosis, which can also lower the cost by decreasing the number of capsules.

First Report of Plum bark necrosis stem pitting-associated virus in Stone Fruit Trees in China.

Plum bark necrosis and stem pitting disease was first observed on a 'Black Beaut' plum (Prunus salicina Lindl.) in the United States in 1986 and later is several other countries. Plum bark necrosis stem pitting-associated virus (PBNSPaV; genus Ampelovirus, family Closteroviridae), the putative causal agent of the disease, infects many stone fruit species and causes decline, gummosis, flattening of scaffold branches, and stem necrotic pits in some diseased trees (1,3). An investigation of the incidence of PBNSPaV on stone fruit trees in China was conducted during 2009 and 2010. Leaf samples were collected from 47 trees, including peach (P. persica L. Batsch), nectarine (P. persica L. var. nucipersica Schneider), plum (P. domestica L.), ornamental plum (P. cerasifera Ehrb), sweet cherry (P. avium L.), and flowering cherry (P. serrulata L.), grown in Hubei, Henan, and Shandong provinces in central and northern China. Most of sampled trees showed trunk gummosis or stem pitting. The presence of PBNSPaV was tested by reverse transcription (RT)-PCR using primer set PBN195F/PBN195R (5'-CTGGTCTTCCTGCTACTCCTT-3'/5'-AAGCCCACAATCTCAGAGCG-3') designed for the detection of the coat protein (CP) gene of the virus. Total RNA was extracted from leaves using a CTAB protocol reported by Li et al. (2). Products of the expected size of 190 bp were amplified from 20 samples, including seven cultivated peach, four ornamental peach, one nectarine, two plum, one ornamental plum, three sweet cherry, and two flowering cherry samples. All trees positive for PBNSPaV showed stem pitting symptoms on the base of the trunk. To further confirm these results, a 590-base region of the heat shock protein 70 homolog (HSP70h) gene was amplified by RT-PCR using primers HSP-P1/HSP-P2 (5'-GGAATTGACTTCGGTACAAC-3'/5'-TCGAAAGTACCACCACCGAA-3'). Amplicons of the expected size were cloned into the vector pMD18-T (TaKaRa, Dalian, China) and sequenced by Genscript Corp. (Nanjing, China). Sequences of 18 PBNSPaV isolates were deposited in GenBank with Accession Nos. JF810177-JF810194. Sequence comparisons showed that the partial HSP70h gene from the Chinese PBNSPaV isolates shared 82.2 to 100% nucleotide (nt) and 94.0 to 100% amino acid (aa) similarities between them and 83.6 to 99.1% nt and 94 to 100% aa similarities with the corresponding region of the other PBNSPaV isolates deposited in GenBank. In July 2010, peach GF305 seedlings were inoculated by side grafting with budwoods from two PBNSPaV-positive ornamental peach plants. In June 2011, grooving symptom was observed on the stems of the seedlings and the virus was detected by RT-PCR. The results further confirmed PBNSPaV infection in China. These results show that PBNSPaV and the associated disease occur in main cultivated and ornamental Prunus species in China. Given the importance and the devastating symptoms of the disease, it is important to prevent virus spread by using virus-tested propagation materials. References: (1) M. Al Rwahnih et al. Arch. Virol. 152:2197, 2007. (2) R. Li et al. J. Virol. Methods 154:48, 2008. (3) D. B. Marini et al. Plant Dis. 86:415, 2002.

First Report of Cherry green ring mottle virus on Cherry and Peach Grown in China.

Cherry green ring mottle virus (CGRMV; a member of the genus Foveavirus in the family Flexiviridae) has a single-stranded, positive-sense RNA genome of approximately 8.4 kb (4). The viral infection on several Prunus spp. has been mainly reported in Japan, New Zealand, and some countries in Africa, Europe, and North America (3). The virus can cause leaf yellowing on sour and tart cherry. Sweet cherry plants are symptomless hosts of the virus. During the growing season of 2010, leaf samples were collected randomly from one ornamental cherry (Prunus serrulata L.) and 26 sweet cherry (P. avium (L.) L.) plants grown in Shangdong and Henan provinces in northern China and 64 peach (P. persica L. Batsch) plants grown in Hubei Province in central China and tested for the presence of CGRMV by reverse transcription (RT)-PCR. Total RNA was extracted from leaves using the CTAB protocol reported by Li et al (2). Primer set, CGRMV1/CGRMV2 (1), was used for the amplification of a 949-bp fragment, which contains the complete CP gene of 807 bp. PCR products with the expected size were identified in one ornamental cherry, seven sweet cherry, and eight peach plants. Although some of sampled plants showed leaf chlorosis, we did not find the specific association between the symptom and CGRMV infection. The obtained PCR products were cloned into the vector pMD18-T (TaKaRa, Dalian, China). Three independent clones from each isolate were sequenced by Genscript Corp., Nanjing, China. Results showed that CP sequences from the Chinese CGRMV isolates shared 87.7 to 99.8% nucleotide and 93.3 to 100% deduced amino acid similarities, and clones intra each isolate shared more than 99% nt similarities. The CP gene sequences of two representative isolates from cherry (YT-Ch-1) and peach (Pe-HB-18) were submitted to GenBank with Accession Nos. HQ539656 and JF810672, respectively. The neighbor-joining phylogenetic trees generated with nucleotide and amino acid sequences of CP genes by Clustal X v1.8 revealed that all Chinese CGRMV isolates fell into two well-resolved clades. Most of the Chinese CGRMV isolates (12 of 16 isolates, including the isolate YT-Ch-1) were grouped in a large clade represented by isolate ITA5 (GenBank Accession No. AF533159). Four isolates from peach (including the isolate Pe-HB-18) clustered into another clade represented by isolate ITA6 (GenBank Accession No. AF533160). In July 2010, peach GF305 seedlings were inoculated by side grafting with budwoods from two CGRMV positive cherry plants. In May 2011, some newly developed leaves from all inoculated plants showed vein yellowing. The CGRMV infection in these inoculated peach GF305 plants was detected by RT-PCR and protein A sandwich-ELISA using antiserum raised against the recombinant CP of CGRMV isolate YT-Ch-1 (unpublished data). These results further confirmed the CGRMV infection on field cherry plants as detected by RT-PCR. To our knowledge, this is the first record of the presence of CGRMV in ornamental and sweet cherry and peach plants in China, which provides valuable information for further evaluating the sanitary status of the virus in sweet cherry and peach orchards in China. References: (1) R. Li and R. Mock. J. Virol. Methods 129:162, 2005. (2) R. Li et al. J. Virol. Methods 154:48, 2008. (3) K. G. Parker et al. USDA. Agric. Handb. No. 437:193, 1976. (4) Y. Zhang et al. J. Gen. Virol. 79:2275, 1998.

A safety assessment of a fowlpox-vectored Mycoplasma gallisepticum vaccine in chickens.

A recombinant fowlpox virus vaccine expressing key protective Mycoplasma gallisepticum antigens could facilitate in the prevention both of fowlpox virus and M. gallisepticum infections. Vectormune FP-MG vaccine, a recombinant fowlpox virus expressing both M. gallisepticum 40k and mgc genes, was assessed for its safety in 8-wk-old specific-pathogen-free White Leghorn chickens. The vaccine virus was serially passaged 5 times by wing-web inoculation. Based on the postinoculation clinical observation, gross pathological examination of air sacs and peritoneum, genetic stability evaluation, virus shedding and tissue distribution detection, horizontal transmission ability determination, and protection against fowlpox virus challenge, the Vectormune FP-MG vaccine possesses a high level of safety.

[Prediction of acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation by the level of galectin-9 in peripheral blood].

Objective: To evaluate possible effects of Gelctin-9 on acute graft versus host disease (aGVHD) development and clinical outcomes in patients before and afer allogeneic hematopoietic stem cell transplantation (allo-HSCT) . Methods: Peripheral blood samples were obtained from 29 patients and 15 healthy volunteers with heparin anticoagulant tubes. Samples were analyzed using ELISA kits to measure the serum concentrations of Galectin-9. Results: Patients developing aGVHD had significantly lower level of Galectin-9 [ (7.96±1.18) µg/L] before allo-HSCT compared with those not developing aGVHD [ (12.37±0.97) µg/L, P<0.001]. And after allo-HSCT, the consentration of Galectin-9 increased markedly in patients developing aGVHD [ (17.78±1.78) µg/L] compared with those not developing aGVHD [ (9.45±0.80) µg/L, P<0.001]. Patients developing 3-4 grade aGVHD had significantly higher level of Galectin-9 [ (23.25±2.59) µg/L] compared with those developing 1-2 grade aGVHD [ (14.37±1.45) µg/L, P=0.008] and those without aGVHD [ (9.45±0.80) µg/L, P<0.001]. The patients with lower level of Galectin-9 after allo-HSCT (<13.61 µg/L) showed more favorable clinical outcomes compared with those with higher level of Galectin-9 (≥13.61 µg/L) . The 3-year overall survival rates were (100.00±6.05) % and (69.23±12.80) %, respectively (P=0.009) . The cumulative incidence of non-relapse mortality was significantly higher in high Galectin-9 group [ (23.08±11.69) %] in comparison with low Gaelctin-9 group [ (0.00±7.39) %] (P=0.023) . There was no significant difference between the two groups in terms of the cumulative incidence of relapse. The cumulative incidence of relapse at 3 years were (8.33±7.98) % and (12.50±8.27) % in high and low Galectin-9 groups, respectively (P=0.708) . Conclusions: The serum concentration of Galectin-9 at the time of engraftment after allo-HSCT may be used as a predictor for the development and severity of aGVHD. Galectin-9 might be considered as a potential new approach to regulate transplant rejection to achieve desirable survival.